met1 DNA Methyltransferase Controls TERT Gene Expression: A New Insight to The Role of Telomerase in Development

M. Zangi, Mohammad Bagher Bagherieh Najjar, M. Golalipour, Mahnaz ghdasi
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引用次数: 4

Abstract

Objective: DNA methylation systems are essential for proper embryo development. Methylation defects lead to developmental abnormalities. Furthermore, changes in telomerase gene expression can affect stability of chromosomes and produces abnormal growth. Therefore, defects in both methylation and telomerase gene expression can lead to developmental abnormalities. We hypothesized that mutation in the methylation systems may induce developmental abnormalities through changing telomerase gene expression. Materials and Methods: In this experimental study, we used Arabidopsis thaliana (At) as a developmental model. DNA was extracted from seedlings leaves. The grown plants were screened using polymerase chain reaction (PCR) reactions. Total RNA was isolated from the mature leaves, stems and flowers of wild type and met1 mutants. For gene expression analysis, cDNA was synthesized and then quantitative reverse transcription PCR (qRT-PCR) was performed. Results: Telomerase gene expression level in homozygous met1 mutant plants showed ~14 fold increase compared to normal plants. Furthermore, TERT expression in met1 heterozygous was~ 2 fold higher than the wild type plants. Conclusion: Our results suggested that TERT is a methyltransferase-regulated gene which may be involved in developmental abnormities causing by mutation in met1 methyltransferase system.
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met1 DNA甲基转移酶控制TERT基因表达:端粒酶在发育中的作用的新见解
目的:DNA甲基化系统对胚胎正常发育至关重要。甲基化缺陷导致发育异常。此外,端粒酶基因表达的改变会影响染色体的稳定性,导致染色体生长异常。因此,甲基化和端粒酶基因表达的缺陷都可能导致发育异常。我们假设甲基化系统的突变可能通过改变端粒酶基因表达诱导发育异常。材料与方法:本实验以拟南芥(Arabidopsis thaliana, At)为发育模型。从幼苗叶片中提取DNA。利用聚合酶链反应(PCR)对植株进行筛选。从野生型和met1突变体的成熟叶、茎和花中分离到总RNA。基因表达分析采用cDNA合成,然后进行定量反转录PCR (qRT-PCR)。结果:端粒酶基因在met1纯合突变株中的表达量比正常植株增加了约14倍。此外,TERT在met1杂合子中的表达量比野生型植株高约2倍。结论:TERT是一个甲基转移酶调控基因,可能参与了met1甲基转移酶系统突变引起的发育异常。
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